Celli Anna, Gratton Enrico
Dermatology Department, University of California San Francisco, San Francisco, CA 94121, USA.
Biochim Biophys Acta. 2010 Jul;1798(7):1368-76. doi: 10.1016/j.bbamem.2009.12.002. Epub 2009 Dec 16.
Scanning-fluctuation correlation spectroscopy was used to detect subresolution organizational fluctuations in the lipid liquid-crystalline phase for single lipid model systems. We used the fluorescent probe Laurdan which is sensitive to the amount of water in the membrane to show that there is a spatial heterogeneity on the scale of few pixels (the size of the pixel is 50 nm). We calculated the pixel variance of the GP function and we found that the variance has a peak at the phase transition for 3 different samples made of pure lipids. The pixel variance has an abrupt change at the phase transition of the membrane and then it slowly decreases at higher temperature. The relatively large variance of the GP indicates that the liquid phase of the membrane is quite heterogeneous even several degrees higher than the phase transition temperature. We interpreted this result as evidence of an underlying microscale structure of the membrane in which water is not uniformly distributed at the micron scale. Imaging of these microstructures shows that the pixels with different GP tend to concentrate in specific domains in the membrane. In the case of single lipid membrane, the statistical and fluctuation analysis of the GP data shows that even such simple lipid systems are capable of generating and maintaining stable structural and organizational heterogeneities.
扫描波动相关光谱法用于检测单个脂质模型系统中脂质液晶相的亚分辨率组织波动。我们使用对膜中水量敏感的荧光探针Laurdan,以表明在几个像素(像素大小为50 nm)的尺度上存在空间异质性。我们计算了GP函数的像素方差,发现对于由纯脂质制成的3个不同样品,该方差在相变处有一个峰值。像素方差在膜的相变处有一个突变,然后在较高温度下缓慢下降。GP相对较大的方差表明,即使比相变温度高几度,膜的液相也相当不均匀。我们将这一结果解释为膜存在潜在微观结构的证据,其中水在微米尺度上分布不均匀。对这些微观结构的成像表明,具有不同GP的像素倾向于集中在膜中的特定区域。在单脂质膜的情况下,对GP数据的统计和波动分析表明,即使是这样简单的脂质系统也能够产生并维持稳定的结构和组织异质性。